关键词: 月季, RcbHLH, 基因克隆, 基因表达, 酵母双杂交

Abstract: The complete cDNA sequence of RcbHLH gene was identified to reveal the underlying anthocyanin synthesis mechanism by homology-based cloning in the‘Slater’s Crimson China’（Rosa chinensis）. The bioinformatic characteristics，tissue specific expression pattern，and interaction of this gene with MYB and WD40 were investigated. These results showed full length open reading frame of RcbHLH gene was 2 112 bp. It encoded 703 amino acids（Accession number：KY783912）. The RcbHLH had a conserved base domain in its N terminal and a conserved Helix-Loop-Helix domain in its C terminal. The results of analyzing protein system evolution showed that RcbHLH had high homology with the strawberry bHLH and other bHLH protein of plants in Rosaceae. qRT-PCR showed RcbHLH gene was mainly expressed in petal. The results of yeast two-hybrid experiments indicated a MYB-bHLH-WD40 （MBW）complex was formed through the interacting of RcbHLH，PavMYB and PavWD40. These results provided basic information to further illuminate the functions of the RcbHLH gene.

Key words: Rosa chinensis, RcbHLH, cloning, gene expression, yeast two-hybrid